Revision 3

#28712

Store at -20C

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Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Applications:
W, W-S, IP, ChIP

Reactivity:
H M R

Sensitivity:
Endogenous

MW (kDa):
79

Source/Isotype:
Rabbit IgG

UniProt ID:
#Q01167

Entrez-Gene Id:
3607

Product Usage Information

For optimal ChIP results, use 10 μL of antibody and 10 μg of chromatin (approximately 4 × 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.
Application Dilution
Western Blotting 1:1000
Simple Western™ 1:10 - 1:50
Immunoprecipitation 1:200
Chromatin IP 1:50

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/mL BSA, 50% glycerol, and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Specificity/Sensitivity

FoxK2 (E7C5H) Rabbit mAb recognizes endogenous levels of total FoxK2 protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ala414 of human FoxK2 protein.

Background

Forkhead box (Fox) proteins are a family of evolutionarily conserved transcription factors defined by the presence of a winged helix DNA-binding domain called a forkhead box (1). In humans, there are over 40 known Fox protein family members, divided into 19 subfamilies, which have evolved to regulate gene transcription in diverse and highly specialized biological contexts throughout development (2). Mutations that disrupt the expression of Fox gene family members have consequently been implicated in a broad array of human disorders, including immunological dysfunction, infertility, speech/language disorders, and cancer (3,4).

FoxK1 and FoxK2 belong to one of the subfamilies of Fox proteins and contain a forkhead-associated (FHA) domain and a conserved forkhead DNA-binding domain (5). FoxK2 was initially identified as a regulator of IL-2 transcription (6). Studies have shown that FoxK2 binds to DNA harboring G/T mismatches and might be involved in DNA mismatch repair (7). FoxK2 also mediates the oncogenic function of adenovirus (Adv) oncoprotein E1A and papillomavirus E6 proteins via association with these proteins (8). Moreover, FoxK2 promotes AP-1-mediated transcriptional regulation by enhancing the recruitment of AP-1 to chromatin (9). FoxK2 is phosphorylated at Ser368 and Ser423 in a CDK1-cyclin B-dependent manner and is involved in cell cycle regulation (10).

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

W: Western Blotting W-S: Simple Western™ IP: Immunoprecipitation ChIP: Chromatin IP

Cross-Reactivity Key

H: Human M: Mouse R: Rat

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

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Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 3

Cell Signaling Technology Logo
Western blot analysis of extracts from NCI-H522, KNRK, and CTLL-2 cells using FoxK2 (E7C5H) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western Blotting Image 1: FoxK2 (E7C5H) Rabbit mAb
Western blot analysis of extracts from various cell lines using FoxK2 (E7C5H) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). Low expression of FoxK2 protein in SR, Hs 675.T, and OVISE cells is consistent with the predicted expression pattern.
Western Blotting Image 2: FoxK2 (E7C5H) Rabbit mAb
Simple WesternTM analysis of lysates (0.1 mg/mL) from CTLL-2 cells using FoxK2 (E7C5H) Rabbit mAb #28712. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the JessTM Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
Western Blotting Image 1: FoxK2 (E7C5H) Rabbit mAb
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 3

Cell Signaling Technology Logo
Immunoprecipitation of FoxK2 protein from NCI-H522 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is FoxK2 (E7C5H) Rabbit mAb. Western blot analysis was performed using FoxK2 (E7C5H) Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as a secondary antibody.
Immunoprecipitation Image 1: FoxK2 (E7C5H) Rabbit mAb
Chromatin immunoprecipitations were performed with cross-linked chromatin from NCI-H522 cells and either FoxK2 (E7C5H) Rabbit mAb or Normal Rabbit IgG #2729, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. The enriched DNA was quantified by real-time PCR, using human BTD promoter primers, human RBM27 promoter primers, human TOP2A promoter primers, and human LIX-AS1 intron 5 primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
Chromatin Immunoprecipitation Image 1: FoxK2 (E7C5H) Rabbit mAb
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.